Department of Anesthesiology, Jinshan Branch Hospital of Shanghai Sixth People's Hospital, Shanghai 201599, P.R. China.
Department of Anesthesiology, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310005, P.R. China.
Mol Med Rep. 2020 May;21(5):2182-2192. doi: 10.3892/mmr.2020.11024. Epub 2020 Mar 12.
Penehyclidine hydrochloride (PHC) suppresses renal ischemia and reperfusion (I/R) injury (IRI); however, the underlying mechanism of action that achieves this function remains largely unknown. The present study aimed to investigate the potential role of autophagy in PHC‑induced suppression of renal IRI, as well as the involvement of cell proliferation and apoptosis. A rat IRI model and a cellular hypoxia/oxygenation (H/R) model were established; PHC, 3‑methyladenine (3‑MA) and rapamycin (Rapa) were administered to the IRI model rats prior to I/R induction and to H/R cells following reperfusion. Serum creatinine was measured using a biochemistry analyzer, whereas aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) expression levels were detected using ELISA kits. Renal tissue injury was evaluated by histological examination. In addition, microtubule‑associated protein light chain 3B (LC3B) expression, autophagosome formation, cell proliferation and apoptosis were detected in the cellular H/R model. The results demonstrated that I/R induced renal injury in IRI model rats, upregulated serum creatinine, ALAT and ASAT expression levels, and increased autophagic processes. In contrast, pretreatment with PHC or Rapa significantly prevented these I/R‑induced changes, whereas the administration of 3‑MA enhanced I/R‑induced injuries through suppressing autophagy. PHC and Rapa increased LC3B and Beclin‑1 expression levels, but decreased sequestome 1 (p62) expression in the cellular H/R model, whereas 3‑MA prevented these PHC‑induced changes. PHC and Rapa promoted proliferation and autophagy in the cellular H/R model; these effects were accompanied by increased expression levels of LC3B and Beclin‑1, and reduced p62 expression levels, whereas these levels were inhibited by 3‑MA. Furthermore, PHC and Rapa inhibited apoptosis in the cellular H/R model through increasing Bcl‑2 expression levels, and suppressing Bax and caspase‑3 expression levels; the opposite effect was induced by 3‑MA. In conclusion, PHC suppressed renal IRI through the induction of autophagy, which in turn promoted proliferation and suppressed apoptosis in renal cells.
盐酸戊乙奎醚（PHC）可抑制肾缺血再灌注（I/R）损伤（IRI）；然而，其发挥此作用的具体机制尚不清楚。本研究旨在探讨自噬在 PHC 诱导的肾 IRI 抑制中的潜在作用，以及细胞增殖和凋亡的参与情况。建立大鼠 IRI 模型和细胞缺氧/复氧（H/R）模型；在 I/R 诱导前给予 PHC、3-甲基腺嘌呤（3-MA）和雷帕霉素（Rapa），在复氧后给予 H/R 细胞。采用生化分析仪检测血清肌酐，ELISA 试剂盒检测天门冬氨酸氨基转移酶（ASAT）和丙氨酸氨基转移酶（ALAT）表达水平。通过组织学检查评估肾组织损伤。此外，在细胞 H/R 模型中检测微管相关蛋白轻链 3B（LC3B）表达、自噬体形成、细胞增殖和凋亡。结果表明，I/R 诱导 IRI 模型大鼠发生肾损伤，上调血清肌酐、ALAT 和 ASAT 表达水平，并增加自噬过程。相反，PHC 或 Rapa 的预处理可显著防止这些 I/R 诱导的变化，而 3-MA 通过抑制自噬增强 I/R 诱导的损伤。PHC 和 Rapa 增加细胞 H/R 模型中 LC3B 和 Beclin-1 的表达水平，但降低自噬溶酶体相关蛋白 1（p62）的表达水平，而 3-MA 则阻止了这些 PHC 诱导的变化。PHC 和 Rapa 促进细胞 H/R 模型中的增殖和自噬；这些作用伴随着 LC3B 和 Beclin-1 的表达水平增加和 p62 表达水平降低，而 3-MA 则抑制了这些水平。此外，PHC 和 Rapa 通过增加 Bcl-2 的表达水平并抑制 Bax 和半胱天冬酶-3 的表达水平抑制细胞 H/R 模型中的凋亡；3-MA 则产生相反的效果。综上所述，PHC 通过诱导自噬抑制肾 IRI，进而促进肾细胞增殖并抑制凋亡。
